Enumeration and Isolation ofcpe-PositiveClostridium perfringensSpores from Feces

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Abstract
A hydrophobic grid membrane filter-colony hybridization (HGMF-CH) method for the enumeration and isolation ofcpegene-carrying (cpe-positive)Clostridium perfringensspores from feces was developed. A 425-bp DNA probe specific for thecpegene was sensitive and specific when tested with bacterial DNA and pure cultures. The enumeration ofcpe-positiveC. perfringensby the HGMF-CH method proved to be as sensitive as nested PCR combined with the most-probable number technique when tested with fecal samples from healthy individuals. With the aid of the HGMF-CH method, positive hybridization signals were detected from two out of seven fecal samples obtained from healthy individuals. Furthermore,cpe-positiveC. perfringenswas successfully isolated from both of these samples. The detection ofcpe-positiveC. perfringensby the HGMF-CH method is dependent on the ratio ofcpe-positiveC. perfringenscolonies to totalC. perfringenscolonies growing on the HGMF-tryptose-sulfite-cycloserine plate.cpe-positiveC. perfringenscould be isolated if the ratio ofcpe-positiveC. perfringensspores to totalC. perfringensspores was 6 × 10−5or higher. The HGMF-CH method provides an aid in the investigation of fecal samples of patients suffering from food poisoning or other diseases caused bycpe-positiveC. perfringens. The method also offers a new approach in the investigation of the epidemiology ofcpe-positiveC. perfringensstrains.

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